TRIBOLIUM INFORMATION BULLETIN
Number 33
July, 1993
Note……………………………………………………………………………………… ii
Acknowledgements …………………………………………………………………….. iii
Research, Teaching and Technical Notes ………………………………………. 53-118
A note on base population parameters in Tribolium castaneum.
G.K. Gau and M.K. Rao …………………..……………………………………. 55
Effects of overcrowding on Tribolium freemani adults and larvae.
E. Lezcano and A. Sokoloff ..…………….……………………………………. 60
Gamma irradiation of the flour beetle, Tribolium anaphe Hinton
(Coleoptera : Tenebrionidae) : Effects of puparial age on the
reproductive ability. Md. Mahbub Hasan and A.M. Saleh Reza ...………... 64
A Preliminary note on the biology of some Tribolium species
(Coleoptera : Tenebrionidae). Mahbub Hasan and B.J. Selman ………….. 70
Insecticidal properties of five plant materials on two strains of
Tribolium castaneum (Herbst), M.A. Malek and R.M. Wilkins ….…………... 79
Reproductive potential of Tribolium castaneum (Herbst)
(Coleoptera : Tenebrionidae) on feed treated with some plant extracts.
Abdul Mannan, S.M. Rahman, Arif Hossain and Ataur Rahman Khan ..….. 90
Effectiveness of caffeine and castor oil as repellent against Tribolium
castaneum. K.A.M.S.H. Mondal and N. Akhtar ……………………………… 95
Intra and interspecific responses of Tribolium castaneum and Tribolium
confusum to conditioned medium. K.A.M.S.H. Mondal ............................. 100
The simulation of sire genetic evaluation using Tribolium castaneum for
the comparison of two statistical methods. Zhang Lao, Wang Xibin,
Zhang Qineng, Wang Yachun and Liu Wei ………………………………... 105
The repair of genetic damage on Tribolium freemani black and
T. freemani – T. castaneum black hybrids through injection of
b-alanine. Page McGraw and Alexander Sokoloff .……………………... 110
Identification of four species of the castaneum species-group of Tribolium
(Coleoptera : Tenebrionidae). Z. Shengfang and L. Yongping ………….. 113
A Succession of habitats in laboratory cultures, A. Sokoloff …………... 117
Notes-Research, Teaching and Technical
* A NOTE ON BASE POPULATION PARAMETERS IN TRIBOLIUM CASTANEUM
G.K. GAUR AND M.K. RAO
DIVISION OF ANIMAL GENETICS
I.V.R.I., IZATNAGAR 243 122 (U.P.)
The present experiment was carried out to study the inheritance of fecundity
traits (egg number and hatchability) and developmental traits (pupation time and
emergence time) in a population in Tribolium castaneum. The population used in
this investigation was established at Population Genetics Laboratory, I.V.R.I.,
Izatnagar, U.P., India by collecting beetles from flour mills in Izatnagar and random
mating them for 10 generations. The culture media consisted of 95 parts whole
wheat flour, 4 parts dried yeast powder and 1 part vitamin mixture (A, B2 and D3).
The cultures were maintained in a B.O.D. incubator at a temperature of 31 + 0.5 and
a relative humidity of 70 + 5%. The larvae were checked for pupation from 10 th day
onwards and the pupae were sexed.
A total number of 50 males and 200 virgin females (at least 6 days old) were
raised from the main stock, and each male was mated to 4 females by putting them
together in a vial with sufficient medium for about 6 days. The eggs produced over a
period of 24 hours by each female were collected and transferred to fresh vials with
fresh medium and incubated. The larvae from each these vials after 10 days of
incubation were counted. The number of larvae expressed as the per cent of the
number of eggs incubated was reckoned as the hatchability per cent. The larvae
were transferred to individual vials. These vials were checked for pupation after 13th
day onwards and later for emergence. Both pupation time and emergence time for
individual progeny were recorded. Altogether 1159 beetles were scored for pupation
time and emergence time, while egg production and hatchability were recorded for
338 beetles. The averages and standard errors for different traits were calculated.
The heritability estimates for different traits and genetic, phenotypic and
environmental correlations among them were also estimated.
The averages, standard errors and coefficients of variation for different traits
are given in Table 1. The averages of pupation time (14.61 days) and emergence
time (19.39 days) were shorter than those reported by Young (1970) and Wade
(1978). The average number of eggs produced over a period of 24 hours (11.62)
was similar to the report of Campo and Rodriguez (1985). Average hatchability
percentage (68.16) was lower than the value reported by Bartlett (1962). The lower
values of pupation time, emergence time and hatchability per cent in the present
study might be due to genetic differences in the strains, daily handling of larvae from
13th day onwards and to a minor extent to the differences in the environmental
conditions. The lower hatchability could also be due to the fact that only those larvae
surviving up to 10th day were reckoned for calculating the hatchability percentage.
The Least squares analysis of variance showed that both pupation time and
emergence time were not influenced by sex. However, females took a slightly longer
time both to pupate and emerge as compared to males.
The heritability estimates for the traits studied are given in Table 2. The
heritability values were higher for pupation time (0.554 to 0.748), emergence time
(0.493 to 0.812) and hatchability percentage (0.580) while it was lower for egg
number (0.176). The estimates of heritability for pupation and emergence time were
slightly higher than those reported by Sokoloff (1977). The higher estimates of
heritability from dam components of variance than those from sire components of
variance revealed that both pupation and emergence time are influenced to a certain
extent, by maternal and dominance effects. Dawson (1965) also observed that 15, 8
and 21 per cent of variation in pupation time could be attributed to additive genetic,
maternal and dominance effects, respectively. The heritability values for egg
production reported by Verma (1977) and Campo and Rodriguez (1985) were higher
than the estimates in the present study showing the variation among different strains
with regard to influence of additive genetic effects.
The correlations between different traits are given in Table 3. Pupation time
had high and positive genetic, phenotypic and environmental correlations with
emergence time. This is expected, as emergence time is the sum of pupation time
and pupal period. The genetic and phenotypic correlations between egg number
and hatchability were very low.
REFERENCES
Bartlett, A.C. 1962. The influence of population density on mortality and body weight
in Tribolium castaneum. Tribolium Information Bulletin, 5 : 22-24.
Campo, J.L. and Rodriguez, M.C. 1985. Experimental comparison of methods for
simultaneous selection of 2 correlated traits in Tribolium castaneum.
Theoretical and Applied Genetics, 71 : 93-100.
Dawson, P.S. 1965.
Estimation of components of phenotypic variance for
developmental rate in Tribolium. Heredity, 20 : 403-17.
Sokoloff, A. 1977. The Biology of Tribolium. Volume 3. Claredon Press, Oxford.
Verma, S.B. 1977. Effect of selection and gamma irradiation on fecundity in
Tribolium castaneum. M.Sc. dissertation, Post Graduate College of Animal
Sciences, I.V.R.I., Izatnagar, U.P., India.
Wade, M.J. 1978. The primary characteristics of Tribolium populations, group
selected for increased and decreased population size. Evolution, 33 : 197985.
Young, A.M. 1970. Predation and abundance in populations of flour beetles.
Ecology, 51 : 602-19.
Table 1 : Averages of different traits in the base population
Traits
Pupation time (days)
Emergence time (days)
Egg number
Hatchability (%)
Number of
Observations
1159
1159
338
338
Mean
S.E.
C.V.
14.61
19.39
11.62
68.16
0.03
0.03
0.27
1.44
6.58
4.83
42.28
38.84
Table 2 : Heritability values for different traits in base population
Traits
Pupation
time
Emergence
time
Egg number
Hatchability
percentage
Sire
components
0.554
S.E.
0.186
Dam
components
0.748
0.493
0.178
0.176
0.580
0.147
0.202
S.E.
0.148
Sire + Dam
components
0.651
S.E.
0.105
0.812
0.156
0.652
0.103
-
-
-
-
Table 3 : Correlations between pupation time and emergence time and egg number
and hatchability in the base population.
Traits
Pupation – Emergence time
Sire component
Dam component
Sire + Dam component
Egg Number – Hatchability
Genetic
0.91 + 0.04
0.99 + 0.00
0.96 + 0.00
0.25 + 0.36
Correlations
Environmental
0.77
0.59
0.83
- 0.03
Phenotypic
0.87 + 0.02
0.04 + 0.05
Lezcano, E., and Sokoloff, A.
Biology Department
California State University
San Bernardino, California 92407
* Effects of overcrowding on Tribolium freemani adults and larvae.
When Tribolium cultures in vials are overcrowded, a series of ecological
changes may take place, eventually leading to a loss of that culture. These
successive changes can be summarized as follows :
1.
There is an increase in moisture. In an air conditioned lab, this moisture
condenses on the inner surface of the walls of a glass container (a vial or
half-pint bottle).
2.
The condensed water droplets begin to combine with flour particles,
forming a glue.
3.
When the proper moisture level is reached, mold begins to develop.
4.
The mold may trap all stages of the flour beetle, resulting in their death.
The dead larvae and adults begin to decay. The survivors are driven to
the surface of the medium.
5.
Some of the adults and larvae, in search of food, may dig tunnels in the
flour.
6.
Decomposition of the dead bodies may result in the release of ammonia
which is toxic to the beetles.
7.
Some of the larvae or adults may be trapped by the fungal mycelia, and
they die in the tunnels. Other larvae may scavenge on the dead bodies of
larvae or adults.
8.
If there are no dead bodies of imagoes, the larvae undergo lysis of their
internal tissues.
9.
If there are dead or dying imagoes, they may release quinones, which may
cause the death of other beetles as well as the fungi.
10.
The flour may turn to a black liquid, or the flour may turn sticky.
11.
Gradually the flour begins to dry, forming a plug which detaches from the
inner walls of the container. As this plug continues to lose moisture,
eventually it forms a solid dry plug.
These changes in the flour are so characteristic that we asked a student to try
to identify the microorganisms in a vial whose contents had turned liquid and black.
A while later she reported that the material did not contain any live micro-organisms
(bacteria, yeasts, or fungi) nor spores of any kind. This was strange, because mold
will grow at least at some point in the destruction of the beetle culture, so that there
should be at least some spores and / or bacteria. On the other hand, if adult beetles
are present in such cultures, it is possible that when they are trapped or when they
die that the quinones in the reservoir of their odoriferous glands are released, and
oxidize everything they come in contact with. This may explain why the attempts at
the culturing of the contents by the student assistant produced negative results.
This report summarizes the experiments carried out to explain the ecological
changes which occurred in these vials.
Materials and Methods. The organisms used in these experiments were last instar
larvae and adults of Tribolium freemani (because at the time they were available in
large numbers). The experiment was set up on 5/21/93.
There were two sets of 1 dram (5 cc) vials, one set containing larvae only and
the other adults only. All of the vials contained 1 gram of unbleached wheat flour.
The larvae and adults were introduced at densities of 25, 50, 75, 100, 150, 200, 250
per gram. There were 10 replicates for the densities 25-100 per gram, and only 5
replicates for the densities 150-250 per gram.
The two sets of vials were introduced into a walk-in incubator maintained at
300c. Humidity was not controlled in the incubator.
Each week the changes in the vials were recorded. If there were microorganisms evident in the vials, preliminary identification was attempted of the
cultures of these organisms. The experiment was terminated about three months
after it was begun.
Results. The complete results will be published elsewhere. We give here an
overview of the results of vials containing larvae and adults separately.
1.
Vials containing larvae. The vials containing 100, 150, 200 and 250
larvae / gram failed to yield any survivors at the end of three months. Those started
with 25, 50 and 75 larvae / gram of flour produced descending numbers of survivors
as density increased. The overall survivor values (adults, pupae and larvae) were
54% for density 25 / gram, 36% of density 50 / gram, and 33% for density 75 / gram.
With one exception in density 75, none of the vials containing 25, 50 or 75 larvae /
vial developed any mycelia, nor did the medium become compact, solid or damp. All
of the remaining densities (100-250 larvae / vial) developed mycelial plugs (with the
exception of 3 replicates in density 100), which became compact and solid. The
larvae died within the medium.
2.
Vials containing adults. Only adults in densities 25 and 50 / gram
survived. In the density of 25 / gram some adults died, and there were some larval
progeny among the survivors. At density 50 / gram there were fewer larvae, and
overall 54% of the adults survived. In the remaining densities (75, 100, 150, 200 and
250).
The medium was granular at the top and compacted at the bottom in all
densities (100-250 for the first two weeks. In the next four weeks the medium
became solid and damp, and some mycelial growth was noted. Some of the
populations were active, but other became sluggish.
By the time two weeks had elapsed, the food level had been reduced and
probably consisted of fecal matter (it had a grayish granular appearance). The
densities of 200 and 250 / gram had become very damp and had a dark brown color.
The beetles were “swimming” through a rather thick medium. Some fungal growth
was visible under 30X magnification.
At the end of a month some of the cultures started drying out. There was no
apparent food source except for some fungal growth around the inside surface of the
vial, but beetles were still active. Two weeks later there was no longer any evidence
of mycelial growth in most vials, and most of the adults were dead. In the density
200 / gram vials, the medium had formed a plug in one vial, and in another vial the
plug was extremely damp and black. In the density 250 vials there were two vials
that had black plugs. This was at the last observation on 7 / 15 / 93.
Discussion. It is clear that as density increased, there was also an increase in
dampness because of the release of moisture by the beetles. This increase in
moisture, generated by the larvae and adults, may have (with few exceptions) a
deadly effect on the larvae because the development of mycelial growth in the
presence of such moisture may trap the larvae. Apparently the larvae which survive
longest under these conditions will be those who have crawled to the surface of the
medium.
Larvae living in the non-damp media were able to survive. It may be noted
that stink glands do not develop until the pupal stage, so that larvae cannot sterilize
the medium against fungi.
The adults were also affected by an increase in moisture at the high densities
as evidenced by the fact that the medium formed a plug. The “plug” is identical in
the larval and adult vials. However, once the plug is formed, the adults can still
break it down and use it as food. (It is noteworthy that in high adult density vials
many of the dead beetles were represented by body fragments, the remains of
scavenging activities of the survivors increased, judging from the number of dead
beetles reduced to fragments.)
The adults in the high density vials apparently did not attempt to release their
quinones. Had they done so, the confined atmosphere in the vial would surely have
changed the air contents to a gas chamber and killed the beetles. When last
examined, the high density vials contained a few live adults and even a few small
larvae.
Md Mahbub Hasan # and A M Saleh Reza
Department of Zoology
University of Rajshahi, Rajshahi
Bangladesh
* Gamma Irradiation of the Flour Beetle, Tribolium anaphe Hinton (Coleoptera :
Tenebrionidae) : Effects of Puparial Age on the Reproductive Ability.
The flour beetle, Tribolium anaphe Hinton is one of the most destructive pest
infesting grains and their products throughout the many countries. In an attempt to
suppress and control insect pest populations infested stored products, several
methods and techniques are being used, e.g. chemical control, temperature
treatment, fumigation and so on. However, because of insecticide resistance,
gamma radiation appears to be a potential alternative to these for insect control in
stored products (Hasan et al. 1989; Navon et al. 1988; Mehta et al., 1990; Sengonca
& Schade and Saxena et al., 1992). Comparative studies of radiation sensitivity
among stored product insect pests are important for radiation control because the
sensitivity varies depending on the stages and ages (Lee & Ducoff, 1983; Williamson
et al., 1985; Ishii et al., 1986 and Ducoff, 1986). Although the irradiation effects on
the various aspects of Tribolium spp. have been reported by several authors (Corks,
1957; Sokoloff, 1961; Yang, 1973; Mickibben de Conconi, 1978; Bonginwar et al.,
1981; Wool, 1982 and Cheng & Ducoff, 1989). However, a literature survey
concerning the radiation effects revealed that a very little is discussed with T. anaphe
sp. specially on the reproductive potential. So, therefore, in the present paper the
effects of gamma radiation on the reproductive ability of T. anaphe emerged from the
various ages irradiated pupae, are described.
The pupae of T. anaphe used in this study were obtained from the laboratory
stock cultures reared in glass jars (20 X 10 cm) on the complete diet described by
Park and Frank (1948). The gamma radiation (Co60) was exposed to the various
ages pupae such as 1-, 2-, 3- and 4-day old and the doses were 0 (control), 1-, 2-, 4and 6-krad. After exposed, the pupae were kept separately in sex and age wise in
an incubator for eclosion. After eclosion, they were paired sexwise and put in a vial
(3, 4 X 1.8 cm) containing whole meal flour for oviposition. The eggs were counted
by sieving the contents at 3-days intervals for 45-days.
---------------------------------------------------------# To whom correspondence should be sent
The percent of reproductive control (PRC) was calculated as follows (Rizvi et al.,
1980) :
(V1-V2)
PRC = ------------- x 100
V1
Where V1 = eggs laid by the control females and V2 = eggs laid by the treated
females. Eggs were also observed for their fertility. Here fertility has been defined
as the percentage of first instar larvae that emerged from an accurately known
number of eggs (Park et. al., 1961). Each treatment was comprised with 15 pairs of
adult in each age and dose for the oviposition. The adults failed to emerge from the
pupae irradiated at 6-krad in all the age groups. A very few eclosed adult was also
observed in 1-day pupae irradiated at 2- and 4-krad doses. So, it was not possible
to continue the experiments at these treatments with the parameters considered.
The sources of radiation used in the present experiments was the gamma
Co60 and 12 krad / hr at 30 cm distance from the middle of the source pencil. The
selection of radiation dose was based on its effectiveness against Tribolium spp.
(Hasan et al., 1989 and Mehta et al., 1990). All the experiments were carried out in
an incubator at 300c throughout the investigations.
The results showed that gamma radiation affects drastically the fecundity and
fertility of T. anaphe adults emerged from the various ages pupae (Fig. 1). The
fecundity was decreased gradually in increasing the doses. The same sequence
was also observed in the fertility. The d-values indicated that there are significant
variation between the corresponding control of each age and dose (Table 1). The
adults emerged from the 1 day old pupae became moribund and died within 4-5 days
of its emergence when exposed at 2- and 4-krad.
Data concerning the sensibility indicated that younger pupae were more
sensibility to gamma radiation than the older pupae. The ratio-sensitivity was also
decreased in increasing ages. The sequence of the susceptibility among the ages
was 1 > 2 > 3 > 4 day old pupae. The number of progeny per female abruptly
declined from 2 krad to 4 krad in all the age groups (fig. 1). The reduction in number
of progeny was great at 4 krad dose level. North and Holt (1970, 1971) reported that
the reduced fecundity in any insects may cause of reduced transfer of active sperms
by treated males to untreated females and to limited production of oocytes in the
developing female.
The results further indicated that the doses used in the experiments did not
show any of the complete sterility either in the male or female which is in agreement
with those of Banham & Cork (1966). Von Borstel (1968) also reported that the
fertility from the irradiated larvae support the general statement that the egg cells are
more sensitive to radiation than sperm and spermatocytes. The magnitude of the
radiation effect on Tribolium is influenced by the dose and age at exposure (Ducoff,
1975). A previous work by Begum et al. (1985) has shown that the sensitiveness of
Callosobruchus analis to certain gamma doses are positively correlated with
increased ages which also supports the result of the present investigation.
Reviewing the present results and the previous works (Lee & Ducoff, 1983;
Williamson et al., 1985; Ishii et al., 1986), it may be concluded that the age plays an
important role in influencing the sensibility of stored product insects to gamma
irradiation.
Age of
pupae
(days)
1
2
3
4
1 krad
d* - values
PRC **
8.14
7.08
6.91
6.26
28.16
24.93
25.74
8.61
2 krad
d – values
PRC
5.41
7.79
5.34
79.49
78.05
54.88
4 krad
d – values
PRC
4.83
7.12
6.14
88.87
83.34
68.52
* d = Normal standard deviate, ** PRC = Percent Reproductive Control
- Adults not survived
Table 1. Showing the values of normal standard deviate and percent reproductive
control of T. anaphe adults emerged from the various ages irradiated
pupae.
Acknowledgement : The authors would like to thank the Chairman, Department of
Zoology, Rajshahi University, Rajshahi, for providing the necessary laboratory
facilities; the Chairman, Bangladesh Atomic Energy Center, for the radiation facilities
and also the Slough Laboratories, MAFF, England, for kindly supplying the
experimental insects.
References
Banham, E.J. and Cork, L.J. 1966. Susceptibility of the confused flour beetle,
Tribolium castaneum (Herbst) to gamma radiation. Pergamon Press, pp. 107111.
Begum, A.; Matin, A.S.M.A.; Badiuzzaman, M. and Seal, D.R. 1985. Studies on the
effects of gamma radiation on the Callosobruchus analis (F.) (Coleoptera :
Bruchidae). Bangladesh j. zool. 13 (1) : 29-35
Bonginwar, D.R.; Palwal-Desal, S.R.; Sudha Rao, V.; Vakil, U.K. and Sreenivasan,
A. 1981. Semi-plot scale studies on wheat disinfestigation gamma radiation.
J. Food Sci. Tech. 18 (6) : 225-231.
Cheng Chi-Hing, C. and Ducoff, H.S. 1989. High-dose mode of death in Tribolium.
Ent. exp. & appl. 51 : 189-197.
Cork, J.M. 1957. Gamma radiation and longevity of the flour beetle. Radiation Res.
7 : 551-557.
Ducoff, H.S. 1975. Form of the increased longevity of Tribolium after x-irradiation.
Expt. Geront. 10 : 189-193.
Ducoff, H.S. 1986. Radiation and longevity enhancement in Tribolium. In : Insect
Ageing, Springer-Veriag, Berlin.
Hasan, M.; Khalequzzaman, M. and Khan, A.R. 1989. Development of Tribolium
anaphe irradiated as larvae of various ages with gamma rays. Ent. exp. &
appl. 53 : 92-94.
Ishii, M.I.; Honda, M. and Sano, M. 1986. Infestivity and development of gamma
irradiated first stages larvae of Angiostrongteus cantonensis. Z. Parasitenkd.
72 : 331-334.
Lee, Y.J. and Ducoff, H.S. 1983. Age and sensitivity to oxygen in the flour beetle
Tribolium confusum. Mech. of Ageing and Development. 22 : 97-103.
Mckibben de C. 1978. A study of the effects of irradiation on the developmental
stages of Tribolium brevicornis. Tribolium Inf. Bull. 15 : 89-95.
Mehta, V.K.; Sethi, G.R. and Grag, A.K. 1990. Development of Tribolium castaneum
(Herbst) larvae after gamma irradiation of eggs. J. Nucl. Agric. Biol. 19 : 5457.
Navon, A; Yatom, S.; Padova, R. and Ross, I. 1988. Gamma irradiation of
Spodoptera littoralis eggs and neonate larvae to eliminate the pest on flowers
for export. Hassadeh 68 : 710-711.
North, D.T. and Holt, G.G. 1970. Population control of Lepidoptera. The genetic and
physiological basis. Manit. Entomol. 4 : 53-69.
North, D.T. and Holt, G.G. 1971. Radiation studies of sperm transfer in relation to
competitiveness and oviposition in the cabbage looper and corn earworm. P.
87-97. In : Application of Induced Sterility for Control of Lepidopterous
Populations. (Proc. Panel, June, 1970). Int. At. Energy Agency, Vienna, 169
p.
Park, T. and Frank, M.B. 1948. The fecundity and development of the flour beetles,
Tribolium confusum and T. castaneum at three constant temperatures.
Ecology 29 : 368-375.
Fig. 1 : Effect of gama irradiation on the number of eggs per female / day (a), mean
fecundity (b) and fertility (c) of T. anaphe treated as various ages pupae. Line on the
top of the bar indicates the absolute values of standard deviation.
Park, T.; Mertz, D.B. and Pestrusewicz, K. 1961. Genetic strains of Tribolium
: their primary characteristics. Physiol. Zool. 34 : 62-80.
Rizvi, S.J.H.; Pandey, S.K.; Mukerjii, D. and Mathur, S.K. 1980.
1,3,7
Trimethylxanthine a new chemosterilant for stored grain pest, Callosobruchus
chinesis L. Z. angew. Ent. 90 : 378-381.
Saxena, B.P.; Sharma P.R.; Thapp, R.P. and Tikku, R. 1992. Temperature induced
sterilization for control of three stored grain beetles. J. stored Prod. Res. 28
(1) : 67-70.
Sengonca, C.V. and Schade, M. 1991. Sterilization of grape vine moth eggs by
ultraviolet rays and their parasitization suitability for Trichogramma semlidis
(Auriv) (Hymenoptera : Trichogrammatidae) J. Appl. Entomol. 111 (4) : 321326.
Sokoloff, A. 1961. Irradiation experiments with Tribolium. Tribolium Inf. Bull. 4 : 2833.
Von Borstel, R.C. 1968. Control of adults pests by the irradiation-of-male method.
In : Isotopes and Radiation Entomology. IAEA, Vienna, pp 331-337.
Williamson, D.L.; Mitchell, S. and Seo, S.T. 1985. Gamma irradiation of the
Mediterranean fruit-fly (Diptera : Tephritidae) : Effects of puparial age under
induced hypoxia on female sterility. Ann. Entomol. Soc. Am. 78 : 101-106.
Wool, D. 1982. Productivity of x-irradiated Tribolium. Tribolium Inf. Bull. 22 : 171175.
Yang, T.C. 1973. Compound eyes as possible detectors of irradiation in flour
beetles. Tribolium Inf. Bull. 16 : 110-112.
Mahbub Hasan1 and B J Selman
Department of Agric and Env Science
University of Newcastle upon Tyne
NE1 7RU, United Kingdom
* A Preliminary Note on the Biology of Some Tribolium Species (Coleoptera :
Tenebrionidae).
Flour beetles of the genus Tribolium include a large number of species
feeding on a variety of stored commodities throughout the world (Sokoloff, 1972).
They are useful experimental tools which have been used in classic studies of the
biology of ageing. They are easy to culture in large numbers and require no
sophisticated equipment for maintenance. However, the biology of this genus varies
greatly from one species to another (Chapman, 1924; Frost et al., 1944; Reynolds,
1945; Cashman, 1951; Magis, 1954a; Khalifa & Badawy, 1955b; de Faria Estacio,
1956; Howe, 1956; Sang, 1959; Naylor, 1964; House, 1965; Sokoloff et al., 1966b;
Gordon, 1968). The culture media is one of the main factors which controls the rate
of development of Tribolium throughout ontogeny (park and Frank, 1948; Magis,
1963; Sokoloff et. al. 1966a). Many sorts of media have been used by entomologists
to maintain Tribolium cultures, though standard media have been established for the
culture of both T. confusum and T. castaneum (Park and Frank, 1948). However,
there is a lack of published data describing food media suitable for the culture of
other Tribolium species. The present paper is an attempt to determine the effect of
specific food media on the biology of some Tribolium species, i.e. T. anaphe Hinton,
T. brevicornis Leconte, T. castaneum Herbst, T. destructor Uyttenb, T. freemani
Hinton in detail.
Beetles of Tribolium species used in the present experiments were originally
received from the Slough Laboratories, MAFF and were cultured using the following
food media :
Species
T. anaphe
T. brevicornis
T. castaneum
T. destructor
T. freemani
a
Food media
wheat feed : fishmeal :
yeast
rolled oats : Wheatfeed :
fishmeal : yeast
Whole meal : yeast
Wheatfeed : yeast
Whole meal flour : yeast
Ratios
8:4:1
20 : 20 : 1 : 1
19 : 1a
12 : 1
11 : 1
- Park and Frank, 1948
To whom correspondence should be sent
1-
The food media were sterilized for six-hours at 1200c and not used until at
least 15 days after sterilization to stabilize moisture content and to equilibrate with
the environment. Each jar was subcultured every three months to remove dead
insects and exhausted medium. The cultures were kept in 3 lb. kilner jars previously
sterilized at 1800c. Some crumpled filter papers were placed inside each jars for
easy movement of the insects and the jars covered at the top with antibacterial filter
papers.
Some beetles of each species were taken from the culture and kept in a Petri
dish for oviposition. The eggs were collected on the following day using the method
of Khan and Selman (1981) and kept in a Petri dish for hatching. After hatching, 200
neonate larvae of each species were collected with a fine camel hair brush and
transferred to the respective food media in a 2 lb kilner jar. After ten days, the larvae
of each species were weighed and their length and headcapsule width measured.
After pupation, the larval periods were recorded. The pupae were sexed using the
exogenital processes of the female (Halstead, 1963) and weighed. After eclosion,
the pupal periods were recorded and the adults weighed. After ten days, the adults
of each species were paired and placed in a glass vial (2.5 x 5 cm) containing the
respective food medium. The eggs were sieved off at 3 day intervals for 30 days
and kept in a Petri dish to record the fertility rate. The length and width of the eggs
were also measured. All the experiments were conducted in an incubator at 270c
and uncontrolled relative humidity.
The data on the biology of Tribolium species cultured with different media are
shown in Figs. 1-4. The maximum 10 day larval weight of 0.41 mg was found in T.
destructor followed by T. anaphe > T. brevicornis > T. freemani > T. castaneum (Fig.
1a). However, these trends changed in the mature stage where the maximum larval
weight was recorded in T. brevicornis followed by T. destructor > T. freemani > T.
anaphe > T. castaneum (Fig. 2a). Khan & Hasan (1990) and Hasan & Khan (1988)
also observed a 4.54 mg mature larval weight in T. anaphe reared at 30 0c. Rich and
Bell (1982) recorded 1.45 mg for 21 day larval weight in T. brevicornis using
standard food media which does not support the present results, however, this
difference may be due to the use of different food media. In the present results, a
maximum larval weight of 16.98 mg was recorded in T. brevicornis (Fig. 2a). Larval
length and headcapsule width also varied significantly both for the 10 day and
mature larvae (Fig. 1b, c; Fig. 2b, c).
Fig 1 : (a) Ten day larval weight, (b) length and (c) headcapsule width of different
Tribolium species (Ta – T. anaphe, Tb – T. brevicornis, Tc – T. castaneum,
Td – T. destructor, Tf – T. freemani) (Line bars indicate the SD)
Fig 2 : (a) Mature larval weight, (b) length and (c) headcapsule width of different
Tribolium species (Ta – T. anaphe, Tb – T. brevicornis, Tc – T. castaneum,
Td – T. destructor, Tf – T. freemani) (Line bars indicate the SD)
Fig 3 : (a) Pupal, (b) adult weight and (c) larval (d) pupal periods of different
Tribolium species (Ta – T. anaphe, Tb – T. brevicornis, Tc – T. castaneum,
Td – T. destructor, Tf – T. freemani) (Line bars indicate the SD)
Fig 4 : (a) Fecundity, (b) fertility, (c) egg laid / female / day, and (d) egg size of
different Tribolium species (Ta – T. anaphe, Tb – T. brevicornis, Tc – T.
castaneum, Td – T. destructor, Tf – T. freemani) (Line bars indicate the SD)
Within species the pupal and adult weight also varied between the sexes and
these corresponded with their larval weight (Fig. 3a, b). The female was heavier
than the male in all stages. These results are in close conformity with the findings of
Nakakita et al. (1981). The maximum larval period of 45.08 days was recorded in T.
freemani followed by T. brevicornis > T. destructor > T. anaphe > T. castaneum (Fig.
3c). Sokoloff (1992) recorded a 41.65 (240c, 50% rh) day larval period for T.
brevicornis. Nakakita et al. (1981), Hasan & Khan (1988), Khan & Hasan (1990) and
Magis (1954) observed 33.8 (300c, 22% rh), 17.91 (300c, 85% rh), 20.06 (300c, 82%
rh) and 34.00 (270c, 70% rh) day larval periods in T. freemani, T. anaphe, T.
castaneum and T. destructor respectively using a different range of culture media.
However, the pupal periods did not follow the same trends as the larval periods. The
maximum pupal period recorded was 11.45 days in T. brevicornis followed by T.
destructor > T. freemani > T. castaneum > T. anaphe (Fig. 3d). Sokoloff (192),
Nakakita et. al. (1981) and Hasan & Khan (1988b) recorded 12.15 (24 0c, 50% rh),
7.3 (300c, 22% rh) and 4.42 (300c, 70% rh) day pupal periods in T. brevicornis, T.
freemani and T. anaphe respectively using different food media. Mathlein (1943)
also noted a 31 day larval and a 9 day pupal period in T. destructor at 28 0c and 7080% rh.
The maximum reproductive potential of 8.60 eggs / female / day was found in
T. anaphe followed by T. freemani > T. brevicornis > T. destructor > T. castaneum
(Fig. 4a). However, Khan (1981) recorded 3.47 eggs laid / female / day in T.
castaneum using a standard food media (Park and Frank, 1948). The highest fertility
was found in T. castaneum and the lowest in T. destructor (Fig. 4b). Howe (1962b)
and Nakakita et al. (1981) found the fertility percentage in T. castaneum and T.
freemani to be 95% (27.50c, 70% rh) and 92% (300c, 22% rh) respectively. The
longest egg was T. brevicornis followed by T. destructor > T. freemani > T.
castaneum > T. anaphe (Fig. 4d). However, the egg width is not proportional to
length where T. destructor > T. freemani > T. brevicornis > T. castaneum > T.
anaphe (Fig. 4d). Brindley (1930), de Fraria Estacio (1956) and Khan (1981) also
found that the egg size of some Tribolium species showed a significant variation
between each species.
Acknowledgements : The authors would like to thank the Head, Dept. of
Agricultural & Environmental Science, Newcastle upon Tyne University, for providing
the necessary laboratory facilities; Slough Laboratories, MAFF, for kindly supplying
the experimental insects and ODA, for funding this project. The first author is also
grateful to Rajshahi University, Bangladesh for granting his study leave.
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* Insecticidal properties of five plant materials on two strains of Tribolium castaneum
(Herbst).
M.A. Malek* and R.M. Wilkins
Department of Agricultural & Environmental Sciences
University of Newcastle Upon Tyne
NE1 7RU, U.K.
ABSTRACT
The powder seed of Annona squamosa Linn. And the ground leaves of Vitex
nequndo Linn. Were found to be toxic to FSS2 and CTC12 eggs of T. castaneum.
The powder seed of A. squamosa and ground twigs of Polygonum hydropiper Linn.
Were toxic to the larvae and pupae of susceptible and malathion resistant strains of
T. castaneum. None of the tested plant materials including Aphanomixis polystacha
Wall and Sapindus mukorosis Gartn in crude powder form were able to kill either of
the adult insects for any of the strains at the highest concentration (1200 ppm).
INTRODUCTION
Tribolium castaneum commonly referred to as the “red flour beetle” is a major pest of
stored products (Hill, 1990). The occurrence of malathion resistance in T.
castaneum has been reported in almost every strains (Dyte & Black, 1972) and thus
incourged the search for insecticidal action in plant products. Five different platn
materials (in crude dried powder form) were evaluated here against eggs, larvae,
pupae and adults of two strains of T. cast i.e. FSS2, susceptible and CTC12
resistant to malathion respectively. The five different plant species were (1) Annona
squamosa (A.s.) Linn., (2) Aphanomixis polystacha (A.p.) Wall., (3) Polygonum
hydropiper (P.h.) Lilnn., (4) Virtex nequndo (V.n.) Linn and (5) Sapindus mukorosis
(S.m.) Gaertn. These plant have been identified and described by Indian Scientists
to have insecticidal properties (Anon. 1948). The plant were chosen for study in the
present experiments because their activity on T. castaneum is unknown and they are
readily available in tropical and sub-tropical countries.
The dried seed powder of A. squamosa has been tested against Callosobruchus
chinensis as a protectant of mung seed (Pandey & Verma, 1977). The dried and
powdered leaves of five plant namely, Nicotiana tabacum, Virtex nequndo,
Polygonum serralatum, Azadirachta indica and Leucas aspera have been used as
protectant of stored wheat aginst the attach of Sitophilus oryzae (Kabir et al., 1984).
The dried seed powder of A. squamosa and A. reticulata have long been used
against head and body lice throughout many tropical countries (Harper et al. 1947).
--------------------------------------------------------------------------* Correspondence should make to the following address :
M.A. Malek, C/O M.A. Wadud, Vill : Rahimpur, Post : Ishurdi, Dist : Pabna,
Bangladesh
The extract of the seed powder of A. squamosa have been examined for contact and
stomach toxicity and ovicidal properties in a variety of insects e.g. contact toxicity
against Aphis fabae, Macrosiphoniclla samborni etc; stomach poison against Plutella
maculipennis larvae, Diatarxia oleracea larvae etc and ovicidal activities against P.
maculipennis, Ephestia kuhniella (Harper et al. 1947).
The greenish mucilaginous juice of Polygonum hydropiper has been used to kill
mosquito larvae (Chopra et al. 1948). The leaves of Virtex nequndo have been
reported to have insecticidal properties and seeds of A. polystacha and seed
pericarp of S. mukorosis have been found to be toxic against insect and fish
respectively (Anon, 1048). Chawal et al. (1992) reported that the CHCl 3 extract of
the defatted seeds of V. nequndo exhibited anti-inflamatory activity and yielded four
triterpenoids. Pericarps of Sapindus mukorosis have been used as a folk medicine
as well as a source of natural surfactant (Wong et al. 1991). Giganenin and 4deoxygiganteus (Annonaceae) which were active in the brine shrimp lethality test
and also significantly sytotoxic to human tumor cells (Fang et al. 1992).
MATERIALS AND METHODS
Seeds of A. squamosa and A. polystacha; twigs of P. hydropiper; leaves of V.
nequndo and seed pericarp of S. mukorosis were dried at 30 0c in an oven for 12
hours and then grounded in an electric blender and sieved through a 250 micrometer
aperture sieve. Five different doses (1.0, 1.5, 2.0, 2.5, 3.0%) in a food medium
(whole meal flour and dry yeast 19 : 1) were prepared (w/w) for each plant material.
Only food medium (5 g) was used for the control. The two strains of T. castaneum
were collected from the stock culture of toxicology laboratories of this University. All
plant materials were collected from Bangladesh after drying under sunshine at 30 0c.
The adults insects were reared in whole meal flour and dry yeast (19 : 1) for
collecting eggs.
One experiment was conducted on each individual plant material to assess the
mortality of eggs, larvae and pupae of the FSS2 and CTC12 strains of T. castaneum
individually. This experiment was conducted initially with fresh eggs and three sets
of data such as egg, larval and pupal mortality were extracted from the single
experiment because five different plant materials were treated with five different
doses for each plant material and there was four replications for each dose on the
two strains of T. castaneum.
Mortality of the eggs of FSS2 and CTC12 strains of T. castaneum
Ten fresh eggs (24 hours old) of both FSS2 and CTC12 strains were taken for each
replication and were placed with treated and untreated medium individually in flat
bottom glass tubes (75 x 19 mm). The mouth of the tubes were then closed by
cotton wool. These glass tubes were then kept in an incubator at 300c, 70% relative
humidity and total darkness. The number of hatched and unhatched eggs were
counted upto 7 days by sieving the treated and untreated medium every 2 days after
treatment. At every observation any 1st instar larvae from each treatment were
transferred into individual glass tubes (50 x 10 mm) for the second set of experiment.
When the eggs were examined under the microscope, it was found that the colour of
unhatched eggs had changed from shine white to grey or yellow. Unhatched eggs
had the eggs which changed their colour were considered as dead eggs. The
number of hatched eggs were determined by counting the number of larvae and
empty egg shells. The percentage mortality of eggs were corrected by Abbott’s
formula (Abbott, 1925) and then subjected to probit analysis (Busvine, 1971).
Mortality of the larvae of FSS2 and CTC12 strains of T. castaneum
First instar larvae of both strains resulting from the above egg mortality experiment.
Larvae were placed individually in flat bottom glass tubes (50 x 10 mm) with freshly
treated and untreated medium (5 g) and the mouths of the tubes were closed by
cotton wool and kept as before. Every three days treated and untreated food
medium were changed with fresh treated and untreated medium to avoid
conditioning. The number of pupae were counted by sieving the food medium
through a 250 micrometer aperture sieve after 20 to 25 days from hatching. The
total number of dead larvae i.e. 1st instar to six instar were determined by subtracting
the number of pupae formed from the number of the 1st instar larvae used.
Mortality of pupae of FSS2 and CTC12 strains of T. castaneum
All the pupae surviving from the larval mortality experiment were transferred
individually to fresh treated and untreated food medium in closed flat bottom glass
rubes (50 x 10 mm) and kept as before. Every three days the food medium was
changed both treated and untreated until adult emergence. The number of newly
emerged adults were counted by sieving the food medium through a 250 micrometer
aperture sieve. The number of dead pupae was determined by subtracting the
number of adults which emerged from the number of pupae used in each replication
of five different doses for each plant material.
RESULTS AND DISCUSSIONS
Mortality of the eggs of FSS2 and CTC12 strains of T. castaneum
The mortality of eggs of both the strains of T. castaneum when treated by five
different plant materials in crude powder form as shown in Figures 1a-e. The LD50
values are compared in figure 1f. The LD50 values, 95% confidence limits for LD50‘s
chi2 and resistance ratios for CTC12 strain with five different plant materials are
shown in Table 1.
Annoa squamosa seed powder was found to be more toxic than the other four plant
materials on eggs of FSS2 and Vitex nequndo was toxic for the mortality of eggs of
CTC12 strain. The chi2 values did not show any significant heterogeneity.
Mortality of the larvae of FSS2 and CTC12 strains of T. castaneum
The probit mortality of the larvae as a whole i.e. from the 1 st instar to six instar of the
two strains of T. castaneum are shown in Figuers 2a-e and LD50 values are
compared in figure 2f. The LD50 values of the larvae, 95% confidence limits for
LD50‘s, chi2 and resistance ratios are shown in Table 1.
A. squamosa seed powder was found to be more toxic than the other four plant
materials for the larvae of FSS2 strain. Polyugonum hydropiper twig powder was
found to be the most toxic to CTC12 larvae. The chi2 values with 3 degrees of
freedom did not show any significant heterogeneity for either of the strains.
Mortality of the pupae of FSS2 and CTC12 strains of T. castaneum
The probit mortality of the pupae of FSS2 and CTC12 strains are shown in figures
3a-e and the LD50 values are compared in figure 3f. The LD50 values for the pupae
with five different plant materials, 95% confidence limits for LD 50‘s, resistance ratios
and chi2 values are shown in Table 1.
A. squamosa seed powder and P. hydropiper twing powder was found to be more
toxic than the other four plant materials for the mortality of the pupae of FSS2 and
CTC12 strain respectively. The chi2 values with 3 degrees of freedom did not show
any significant heterogeneity for either of the strains of T. castaneum.
The petroleum either insoluble extract from A. reticulate seed has shown some effect
against the eggs of Plutella maculipennis at the highest concentration (1.6 and 0.8%
w/v) (Harper et al. 1947). Crosby (1971) reported that Annona extractives had little
or no ovicidal activity. A. squamosa seed powder when mixed with mung bean
(Vigna radiata L.) at the rate of 0.5 to 2.0 parts per 100 parts of seeds protected the
beans against Callosobruchus maculates infestation for 100 days (Pandey & Verma,
1977). All these previous results are more or less similar with the results of the
present study. Tobacco (Nicotiana tabacum L.) leaf powder significantly affected
egg development. 14% of the fertile eggs were found dead 30 days after infestation
by Caryedon seratus (Delobel & Malonga, 1987). Kawazu et al. (1989) reported that
a new compound from A. squamosa seed powder i.e. neoannonin had ovicidal
properties against the eggs of Drosophila melanogaster. Rajapakse (1990) reported
that the peel of Citrus crematofolia significantly reduced the percent of egg hatch at
0.20 g / 50 seeds.
The active compounds such as drimane, warburganal, muzigadial, ugandensidial
and polygodial were isolated from the leaves, seeds of P. hydropiper (Jansen and
Groot, 1991). Warburganal and muzigadial inhibited the feeding of the larvae of the
two species of African armyworm, the monophagus Spodoptera exempta and the
polyphagus S. littoralis at a concentration of 0.1 ppm.
Polygodial and ugandensidial were also antifeedants for these insects but less
active. The drimane was active against S. frugiperda, Heliothis armigera and H.
virescens. Polygodial was active against diamond moth larvae down to 0.1% and it
is inhibited food intake by fifth instar larvae of Pieris brassicae at a concentration of
200 ppm (Jansen and Groot, 1991). Callus and suspension cultures of P. hydropiper
accumulated the insect antifeedant compound i.e. polygodial (Banthorpe et al. 1989).
Table 1. The LD50‘s, 95% confidence limits for LD50 values, Chi2 with 3 degrees of
freedom and resistance ratio for the mortality of 3 various stages of the two strains of
T. castaneum treated with five different plant materials.
Plant
A.s.
A.p.
P.h.
V.n.
Stages
of
insect
Strains
LD50
Eggs
1.92
Larvae
1.90
Pupae
2.66
Eggs
2.50
Larvae
3.40 *
Pupae
2.70
Eggs
3.10 *
Larvae
3.80 *
Pupae
3.28 *
Eggs
3.81 *
Larvae
3.71 *
FSS2
95%
CFL
1.49 –
2.52
1.74 –
2.18
1.67 –
4.29
1.96 –
3.22
2.24 –
5.06
2.08 –
3.50
2.26 –
4.13
2.59 –
3.97
2.37 –
4.47
2.37 –
5.83
2.48 –
4.99
Chi2
LD50
1.20
6.70 *
1.96
3.66 *
0.61
4.10 *
2.15
4.80 *
0.14
5.57 *
1.46
7.65 *
0.61
6.50 *
0.41
3.27 *
0.62
3.68 *
0.71
4.40 *
0.59
5.80 *
CTC12
95%
CFL
2.63 –
17.87
2.47 –
5.18
2.55 –
6.37
2.59 –
8.83
2.70 –
10.33
2.08 –
26.39
2.74 –
15.00
2.41 –
4.32
2.39 –
5.79
2.68 –
11.52
2.52 –
13.01
R.R.
Chi2
0.29
3.50
2.23
5.18
0.68
1.54
0.21
1.92
0.31
1.64
0.31
2.83
0.45
2.09
0.08
0.86
0.50
1.12
0.14
1.15
0.05
1.56
S.m.
Pupae
3.23 *
Eggs
4.65 *
Larvae
2.55
Pupae
2.82
2.26 –
4.66
2.79 –
6.41
2.18 –
3.14
2.26 –
3.52
0.27
3.94 *
4.21
6.15 *
5.10
5.23 *
4.39
7.11 *
2.62 –
5.28
2.97 –
11.52
2.84 –
8.38
2.50 –
8.52
2.03
1.22
0.27
1.32
0.76
2.05
1.55
2.52
A.s. = A. squamosa, A.p. = Aphanomixis polystacha, P.h. = Polygonum hydropiper,
V.n. = Vitex nequndo, S.m. = Sapindus mukorosis, R.R. = Resistance ratio (LD 50 for
resistance strains / LD50 for susceptible strain), * = LD50 (%) extrapolated, CFL =
confidence limits.
A. reticulate seed extract showed toxicity to Plutella maculipennis larvae at higher
concentration. The leaves of chinaberry caused mortality in the larval stages of corn
earmworm, Heliothis zea and fall armyworm, Spodoptera frugiperda when it was
incorporated into a meridic diet or applied to corn seedling grown in green house
(McMillan et al. 1969). The seed extract of A. squamosa and Azadirachta indica
have gustatory activity against the larvae of Sitophilus oryzae and T. castaneum
(Qadri, 1973). Neem (Azadirachta indica) seed and leaf extract have produced 33%
larval mortality of T. castaneum at 8.0% treatment (Pereira & Wohlgemuth, 1982).
From this result it can be said that the seed and leaves extract of neem is less toxic
than A. squamosa seed powder on the larval mortality of T. castaneum. The neem
(A. indica) seed extract incorporated into an artificial diet at 0.02, 0.2 and 2.0% (w/w)
has induced mortality in all larval stages of Trichoplusia ni and Spodoptera exiuga
(Prabhakar et al. 1986). Tobacco leaf powder produced 30 to 56% mortality of T.
confusum larvae at 10 x 103 and 5 x 103 ppm after 20 days of the treatments
(Khalequzzaman et al. 1988). Neoannonin from A. squamosa seed oil fraction had
larvicidal activity at 125 to 140 µg / 2 g of diet against Drosophila melanogaster
(Kawasu et al. 1989). Neoannonin was a pure compound, therefore its toxicity was
more than seed powder of A. squamosa in crude formed on the eggs mortality of
different insects.
The flour of Lathyrus sativus L. produced a significant reduction in pupal and adult
emergence of T. anaphe (Khan & Hasan, 1988). Powder of Piper nigrum
significantly reduced the oviposition and adult emergence in Callosobruchus
maculates (Rajapakes, 1990). Both peppers (ground black and red) reduced the
adult emergence of Callosobruchus chinensis L. at high concentration (1200 ppm)
(Morallo-Rejesus et al. 1990). Nicotine reduced the survival and pupal weight and
prolonged the development of Heliothis virescens F. (Gunasena et al. 1990).
None of the above described plant materials in crude powder form were able to kill
any adult beetles of FSS2 and CTC12 strains of T. castaneum. From this result it
can be said that the crude plant materials were unable to penetrate into the cuticle of
the beetle or unable to produce a toxic effect on the exocuticle of the beetle which is
harder than other stages of the beetle i.e. egg, larvae and pupae. The hardness of
the cuticle of the beetle is due to the presence of a horney substance called selerotin
in the exocuticle. E1-Nahal et al. (1989) reported that no toxic effect could be found
in the adults of Rhizopertha dominica and T. confusum when treated with the
essential oil of Acorus calamus L.
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El-Nahal, A.K.M.; Schmidt, G.H. & Risha, E.M. (1989). Vapour of Acorus calamus oil
– a space treatment for stored product insects. J. Stored Prod. Res. 25 (4) :
211-216.
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Gunasena, G.H.; Vinson, S.B. and Williams, H.J. (1990). Effects of Nicotine on
growth, development and survival of the Tobacco budworm (Lepidoptera :
Noctidae) and the parasitoid, Campoletis sonorensis (Hymenoptera :
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Harper, S.H.; Potter, C. and Gillham, E.M.(1947). Annona species as insecticides.
Ann. Appl. Biol. 34 : 104-112.
Jansen, B.J.M. and Groot, A.D.E. (1991). The occurrence and biological activity of
drimane sesquiterpenoids. Nat. prod. Reports. 83 (3) : 309-316.
Kabir, K.H.; Mia, M.D. & Ahmed, S.U. (1984). Potential uses of some indigenous
plant materials as repellent against rice weevil, Sitophilus oryzae L. on stored
wheat. Univ. J. Zool. Rajshahi Univ. Bangladesh 3 : 41-44.
Kawazu, K.; Alcantara, J.P. & Kobayashi, A. (1989). Isolation and structure of
Neoannonin, a novel insecticidal compound from the seeds of Annona
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Khalequzzaman, M.; Mondal, K.A.M.S.H. & Haque, M.S. (1988). Combined actions
of pirimiphos methyl and tobacco leaf (Nicotiana tabacum L.) powder on the
larval mortality of the confused flour beetle, Tribolium confusum Duval. Tri.
Inf. Bull. 28 : 63-66.
Khan, A.R. & Hasan, M. (1988). Growth on the red flour beetle, Tribolium
castaneum H. (Coleoptera : Tenebrionidae) on Lathyrus sativus L. flour.
Univ. J. Zool. Raj. Univ. Bangladesh. 7 : 1-6
McMillan, W.W.; Bowman, M.C.; Burton, R.L.; Starks, K.J. & Wiseman, B.R. (1969).
Extract of chinaberry leaf as a feeding deterrent and growth retardant for
larvae of the cornworm and fallworm. J. Econ. Ent. 62 (3) : 708-710
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actions of several plants to Callosobruchus chinesis L.. In : Fujii, K. (Ed)
Bruchids and Legumes : Economics, Ecology and Co-evolution. Kulwer
Academic Publisher, Netherland pp. 97-100.
Pandey, G.P. & Verma, B.K. (1977). Annona squamosa (custard apple) seed
powder as a protectant of mung against pulse beetle, Callosocbuchus
maculates F. Bull. Grain Tech. 15 (2) : 100-104
Pareira, J. & Wohlgemuth, R. (1982). Neem (Azadirachta indica A.) of west African
origin as a protectant of stored maize. Z. Angew. Entomol. 94 (2) : 208-214
Prabhakar, N.; Coudriat, D.L.; Kishba, A.N. & Meyerdirk, D.E. (1986). Laboratory
evaluation of neem seed extract against the larvae of the cabbage looper and
beet armyworm (Lepidoptera : Noctuidae). J. Econ. Ent. 79 : 39-41.
Qadri, S.S.H. (1973). Insect suitable for the microbioassy of insecticides residues.
Pesticides (India) 7 (2) : 22-23.
Rajapakes, R.H.S. (1990). Effects of five botanicals as protectans of greengram
against the pulse beetle, Callosobruchus maculatus In : Fujii (Ed) Bruchids
and legumes : Economica, Ecology and Co-evolution. Kulwer Academic
Publisher, Netherland. PP. 85-90
Wong, W.H.; Kasai, R.; Choshi, W.; Nakagawa Y.; Mizutani, K.; Ohtani, K. and
Tanaka, O. (1991). Acyclic sesquiterpene oligoglycerisides from pericarps of
Sapindus delavayi. Phytochemistry 30 (8) : 2699-2702.
Abdul Mannan, S.M. Rahman, Arif Hossain
and Ataur Rahman Khan1
Department of Zoology
Rajshahi University
RAJSHAHI – 6205
BANGLADESH
* Reproductive Potential of Tribolium castaneum (Herbst)
Tenebrionidae) on Feed Treated with Some Plant Extracts.
(Coleoptera
:
The rust-red flour beetle, Tribolium castaneum (Herbst) is a major pest of
several stored products throughout the world. The magnitude of hazards of chemical
pesticides has intensified the search for safer compounds. Safe and inexpensive
methods coupled with simple application techniques are needed. Botanicals are an
attractive alternative to chemical pesticides in being host-specific and biodegradable.
Further, because of their low mammalian toxicity these are particularly valued for
application against pests of fodders, fruits, vegetables and stored grains where
safety is of prime importance.
Fecundity and fertility are two important factors having a direct bearing on
insect populations. The present investigation reports the effect of leaf extracts of
Bishkanthali (Polygonum hydropiper), Neem (Asadirachta indica) and Nishinda
(Vitex negundo) on these parameters of T. castaneum, if any.
Adult T. castaneum were collected from the stock maintained at the Ecology
Laboratory, Department of Zoology, Rajshahi University and were reared on wheat
flour.
The leaf extracts were prepared by cold process in methanol as the solvent
(Worsley 1934). Powders were prepared by pulverization of calcium carbonate in
the extracts of Bishkanthali, Neem and Nishinda leaves in ratios of leaf extracts of
calcium carbonate in the order 1 : 2.23, 1 : 2.23 and 1 :0.75 respectively. A standard
mixture of whole wheat flour with powdered yeast (19 : 1) was used as the
experimental feed. The doses viz. 1.0 x 10 4, 2.0 x 104, 3.0 x 104, 4.0 x 104, 5.0 x 104,
6.0 x 104, 7.0 x 104 and 8.0 x 104, were prepared by mixing the required amounts of
the powders with the food medium.
Neonate T. castaneum larvae were reared on treated food up to pupation.
Similar batches of control were maintained on untreated food. The resultant pupae
were sexed (Halstead 1963) and kept in separate petri dishes for adult eclosion.
After their emergence, adults of opposite sexes were kept in individual glass vials
(3.5 x 1.8cm) containing food. For each dose 10 females were considered. Eggs
were counted after sieving the contents of vials at 3-day intervals for a period of 30
days. Eggs were also observed for their fertility. The experiments were conducted at
30 + 0.20c without any humidity control.
The effect of treatments was to reduce the fecundity of T. castaneum in order
Bishkanthali > Neem > Nishinda (Table 1). The fertility of the female beetles was
also significantly reduced by treatments with the experimental leaf extracts in the
order Nishinda > Bishkanthali > Neem (Table 2).
------------------------------------------------------1To whom correspondence should be addressed
Few works have been conducted on the effect of plant extracts on Tribolium
spp. (Paul et al. 1965, Qadri 1973, Pereira and Wohlgemuth 1982, Golob et al. 1982,
Khanam et al. 1990a, b, Hossain 1990, Jahan et al. 1991, Khalequzzaman and Islam
1992a, b). Botanicals possess a great potential as pesticidal agents.
According to Dick (1937), T. castaneum belongs to the second group of the
four types of egglaying observed in Coleoptera, producing eggs steadily over a long
period of time. The intrinsic rate of increase of T. castaneum is primarily determined
by oviposition early in life (Howe 1962). The results reported in the present
investigation are very much promising regarding the management of this
cosmopolitan pest.
Acknowledgements :
The authors are extremely grateful to the Director, BCSIR Laboratories,
Rajshahi, for supplying the experimental plants and the Chairman, Department of
Zoology, Rajshahi University, for providing necessary laboratory facilities. They
thank Mr. Saiful Islam Faruki, a Ph.D. Fellow, Institute of Biological Sciences,
Rajshahi University for his help.
Literature cited
Dick J. 1937. Oviposition in certain Coleoptera. Ann. App. Biol. 24 : 762-796.
Golob, P., Mwambula, J., Mhango, V. and Ngulube, F.F. 1982. The use of locally
available materials as perfectants of maize grain against insect infestation
during storage in Malawi. J. Stored Prod. Res. 18 : 67-74.
Halstead, D.G.H. 1963. External sex differences in stored-products Coleoptera.
Bull. Entomol. Res. 54 : 119-134.
Hossain, A. 1990. Toxicity of some indigenous plant extract on the rust-red flour
beetle, Tribolium castaneum (Herbst) (Coleoptera : Tenebrionidae). M.Sc.
thesis, Dept. of Zoology, Rajshahi Univ. 112 pp.
Howe, R.W. 1962. The effect of temperature and humidity on the oviposition rate of
Tribolium castaneum (Hbst.) (Coleoptera : Tenebrionidae). Bull. Entomol.
Res. 53 : 301-310.
Jahan,S., Mannan, A., Khan A.R. and Karmaker, P. 1991. Insecticidal effect of
Akanda (Calotropis procera) on Tribolium confusum Duval (Coleoptera :
Tenebrionidae). Bangladesh J. Zool. 19 (2) : 261-262.
Khalequzzaman, M. and Islam, M.N. 1992a. Pesticidal action in Dhutura, Datura
metal Linn. Leaf extracts on Tribolium castaneum (Herbst). Bangladesh j.
zool. 20 (2) : 223-229.
Khalequzzaman, M. and Islam, M.N. 1992b. Synergism of Datura metal Linn. Leaf
and seed extractions with methacrifos on Tribolium castaneum (Herbst).
Tribolium Inf. Bull. 32 : 72-77.
Khanam, L.A.M., Talukder, D., Khan A.R. and Rahman, S.M. 1990a. Insecticidal
properties of Royna, Aphanamixis polystachya Wall. (Parker) (Meliaceae)
against Tribolium confusum Duval. J. Asist. Soc. Bangladesh, Sci. 16 (2) :
71-74.
Khanam, L.A.M., Talukder, D. and Khan A.R. 1990b. Insecticidal property of some
indigenous plants against Tribolium confusum Duval (Coleoptera :
Tenebrionidae). Bangladesh j. zool. 18 (2) : 253-256.
Paul, C.F., Agrawal, P.N. and Ausat, A. 1965. Toxicity of solvent extract of Acorus
calamus Linn. to some grain pests and termites. Indian J. Ent. 27 (1) : 114117.
Pereira, J. and Wohlgemuth, R. 1982. Neem (Azadirachta indica A. Juss) of west
African origin as a protectant of stored maize. Z. angew. Entomol. 94 (2) :
208-214.
Qadri, S.S.H. 1973. Some new indigenous plant repellent for storage pests.
Pesticides 7(2) : 18-19.
Worsley, R.R. 1934. The insecticidal properties of some East African Plants. Ann.
Appl. Biol. 21 : 645-669.
Table 1
Fecundity of T. castaneum females reared on treated food medium
Doses
(ppm)
0(Control)
Bishkanthali
Mean + S.D. Range
61.80 + 7.71
54-77
1.0 x 104
49-70
2.0 x 104
58.00 +
11.30
54.90 + 6.08
3.0 x 104
54.20 + 5.85
42-61
4.0 x 104
5.0 x 104
45-61
33-64
7.0 x 104
53.60 + 5.54
51.80 +
10.62
50.10 +
11.98
46.00 + 7.92
8.0 x 104
44.70 + 7.12
32-56
6.0 x 104
47-67
33-76
34-59
Neem
Mean + S.D.
74.10 +
19.20
60.40 +
16.79
56.40 +
17.28
52.20 +
26.47
51.20 + 9.25
47.10 + 9.13
43.70 +
18.41
38.50 +
16.44
36.70 +
12.81
Range
37-104
35-80
Nishinda
Mean + S.D. Range
65.00 + 9.62 49-81
30-71
26-76
47.90 +
11.64
47.50 + 4.60
13-96
46.90 + 7.52
39-59
39-67
32-61
41.70 + 6.13
36.90 + 7.70
32-48
24-48
16-75
34.90 + 5.70
29-45
26-72
33.50 + 5.99
25-41
21-51
25.70 + 2.67
22-29
42-55
K.A.M.S.H. Mondal and N. Akhtar
Department of Zoology
Rajshahi University
Rajshahi – 6205
Bangladesh
* EFFECTIVENESS OF CAFFEINE AND CASTOR OIL AS REPELLENT AGAINST
TRIBOLIUM CASTANEUM
ABSTRACT
In a food preference chamber, the effectiveness of caffeine and castor oil as
repellents against Tribolium castaneum was studied. Both caffeine and castor oil
were found to be repellent to adults and larvae of T. castaneum. Castor oil was
more effective than caffeine. There was a significant difference (P < 0.05) in
response between sexes of adults – males being more responsive than females of
castor oil. Both early and late larval instars were also more responsive than other
instars.
INTRODUCTION
Tribolium castaneum Herbst is a major pest of stored products and is cosmopolitan
in distribution (Good, 1933, Sokoloff, 1972, 1974). Both adults and larvae are able o
exploit a wide variety of stored commodities (Ziegler, 1977).
In many countries locally available plant materials are widely used as protectants of
stored products against insect pests. The effectiveness of many plant derivatives for
use against grain pests has been reviewed by Jacobson (1958, 1975, 1990). The
insect repellent and antifeedant properties of neem (Azadirachta indica), tobacco
(Nicotiana tabaccum), Nishinda (Vitex negundo), Bishkathali (Polygonum
serrulatum), Dhandakalas (Leucas spera), turmeric (Curcuma longa) have been
reported against stored product pests viz. Trogoderma granarium (Jotwani and
Sircar, 1965), Rhyzopertha dominica (Pereira and Wohlgemuth, 1982), Sitophilus
oryzae (Ahmed et al. 1980, Parveen and Mondal, 1992), Tribolium confusum
(Mondal an Begum, 1991) and Sitotroga cerealella (Abraham et al. 1973). Moreover
different plant oils viz. soybean oil, peanut oil, cottonseed oil, maize oil, rice bean oil,
groundnut oil and citrus oil have also been reported as protectants of stored products
against insect pests (Pandey et al. 1977, 1981; Shaaya and Ikan, 1980; Yadara,
1971; Su et al. 1972a, b; Singh et al., 1978; Schoonhoven, 1978; Qi and Burkholder,
1981; Messina and Renwick, 1983; and Jilani et al. 1988).
In the previous experiment both caffeine and castor oil were found to be toxic to
adults and larva of T. castaneum (Mondal and Akhtar, 1992). However, there is no
information concerning the effectiveness of caffeine and castor oil as either repellent
or attractant against stored product-pests. This led to the present work. Caffeine
(1,3,7 – Trimethylxanthine) – an important compound of coffee (Coffea arabica) and
tea (Cornellia sinesis) is the most important purine and mutagenic agent. It is also
an alkaloid and is responsible for stimulating action on central nervous system
(Noller, 1958; Rakoff and Rose, 196; Cordell, 1981). Castor oil from the seed of the
castor bean plant (Ricinus communis) yields fat acid characterized by the high
concentration of the hydroxyl unsaturated acid, picinoleic acid (DePuy and Rinehart,
1975).
MATERIALS AND METHODS
The experiment was conducted in a ‘choice chamber’ consisting of a plastic petridish
(5 cm diameter) divided into two equal halves by a mark on the outer surface. Each
half was loaded with approximately 1 g of flour, either fresh or treated with caffeine
and castor oil. Ten adults of either sex or larvae were introduced at the middle of the
dish and was covered with a lid, thus providing an option for the test insects to select
either the fresh medium or treated medium (Mondal, 1983). The petridish was kept
in an incubator at 300c. The number of test insects on each half of the petridish was
recorded after 24 hours by sieving the flour medium. The tests were replicated five
times of each treatment and conducted with both larvae and adults separately.
Larve used in the experiment were collected from a T. castaneum larval culture on
6th, 9th, 12th and 18th day from hatching which correspond to third, fourth, fifth and
sixth instars in control respectively (Mondal, 1984). Newly hatched (first instar) and
second instar larvae were not used in the experiment because of their small size.
Sexes of adults were determined in the pupal stage (Halstead, 1963) and adult of
both sexes aged between 10-20 days were used in the experiment. In each test
fresh larvae or adults were used. Flour media treated with either caffeine at the
concentrations of 250, 500 and 1000 ppm or castor oil at the concentrations of 100,
200 and 300 ppm were used in the experiments.
RESULTS AND DISCUSION
The results of the experiments and statistical analyses are shown in Table 1 and 2.
T. castaneum adults and larvae were repelled by both caffeine and castor oil. There
were variable response of adults to different concentrations of caffeine. Adults of
both sexes were either attracted or showed no response to low concentrations (250
and 500 ppm) of caffeine. Only males were repelled by caffeine at high
concentration (1000 ppm). Castor oil was found to be more repellent than caffeine.
There was a significant difference (P < 0.05) in response between sexes of adults –
males being more responsive than females. Similarly a higher proportion of third
and sixth instar larvae were repelled by both caffeine and castor oil suggesting that
both early and late larval instars are more responsive than other instars. No larvae
of all instars were found in medium treated with castor oil at a concentration of 300
ppm indicating the medium highly repellent (table 2).
The present result is similar to those of the previous workers who reported the
repellent action of various plant materials against different species of stored product
insect pests (Jotwani and Sircar, 1965; Abraham et al., 1973; Jilani and Malik, 1973;
Qadri, 1973, Ahmed et al. 1980; Golob et al. 1982; Kabir et al., 1984; Mia et al.,
1985; Jilani et al., 1988; Mondal and Begum, 1991). The repellent action of castor
oil is also similar to the findings of Pandey et al. (1977, 1981), Shaaya and Ikan
(1980), Yadara (1971), Su et al. (1972a, b), Singh et al. (1978), Schoonhoven
(1978), Qi and Burkholder (1981), Messina and Renwick (1983), Jilani et al. (1988)
and Parveen and Mondal (1992) who reported soybean oil, peanut oil, cottonseed
oil, maize oil, rice bean oil, groundnut oil, citrus oil, turmeric oil, sweetflag oil and
neem oil repellent against different stored product pests.
The repellent action of both caffeine and castor oil indicates their possible use in the
control of Tribolium in warehouses as repellents (Dethier, 1956). Bags or containers
treated with repellent caffeine or castor oil may prevent Tribolium adults and larvae
from attacking and infesting the food.
Although the post harvest losses of food grains are commonly protected by
insecticides or fumigation, such practices pose human health risks, environmental
hazards and pesticide-resistant pest strains. However, these problems can be
overcome if the compounds of low mammalian toxicity without environmental
hazards are used. Plant derivations that traditionally have been used as grain
protectants in developing countries as repellents may serve this purpose. As
repellent they may offer protection and comfort without disturbing any ecosystem
because repellents are considered safe as they minimize pesticide uses and
environmental hazards (Jilani et al., 1988). In recent years there has been a
resurgence of world-wide interest in natural plant materials as agents for insect pest
control (Jacobson, 1990). Botanicals possess low mammalian toxicity without
environmental hazards. Thus, the use of both caffeine and castor oil as repellent
against Tribolium may probe to be important from the integrated pest management
point of view.
Table 1. Number and Percentage of T. castaneum adults and larvae in medium
treated with caffeine
Conc. (ppm)
250
500
1000
Life stage
(adults = sex;
larvae = age in
days)
Male
Female
6
9
12
16
Male
Female
6
9
12
16
Male
Female
6
9
12
Distribution of adults / larvae
Total nos.
% Total
X2 (ldf.)
32
36
14
24
26
37
34
20
07
24
28
38
14
27
05
17
23
64
72
28
48
52
74
68
40
14
48
56
76
28
54
10
34
46
3.92 *
9.68 **
9.68 **
0.08 N.S.
0.08 N.S.
11.52 ***
6.48 *
2.00 N.S.
25.92 ***
0.08 N.S.
0.72 N.S.
13.72 ***
9.68 **
0.32 N.S.
32.00 ***
5.12 *
0.32 N.S.
16
18
36
3.92 *
Notes-Research, Teaching and Technical
Table 2. Number and Percentage of T. castaneum adults and larvae in medium
treated with castor oil
Conc. (ppm)
100
200
300
Life stage
(adults = sex;
larvae = age in
days)
Male
Female
6
9
12
16
Male
Female
6
9
12
16
Male
Female
6
9
12
16
Distribution of adults / larvae
Total nos.
% Total
X2 (1df)
02
09
09
10
03
02
02
09
03
03
03
06
-
4
18
18
20
6
4
4
18
6
6
6
12
-
42.32 *
20.48 *
20.48 *
18.00 *
38.72 *
42.32 *
42.32 *
20.48 *
38.72 *
38.72 *
38.72 *
28.88 *
-
Five replicates per test, each replicate consisting of 10 test insects (N = 50)
NS – Not significant, P > 0.05; * Significant, P < 0.05; ** Significant, P < 0.01; ***
Significant, P < 0.001
ACKNOWLEDGEMENTS
The authors are grateful to Professor Abdus Salam Bhuiyan, Former Chairman,
Department of Zoology, Rajshahi University for providing facilities to conduct the
experiments. Grateful thanks are due to Professor M.A. Salam for kindly supplying
the experimental sample of caffeine.
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K.A.M.S.H. Mondal
Department of Zoology
Rajshahi University
Rajshahi 6205
Bangladesh
* INTRA AND INTERSPECIFIC RESPONSES OF TRIBOLIUM CASTANEUM AND
TRIBOLIUM CONFUSUM TO CONDITIONED MEDIUM
ABSTRACT
Both intra and interspecific response of adults and larvae to conditioned medium
were studied in Tribolium castaneum Herbst and Tribolium confusum Duval. Flour
conditioned by both T. castaneum and T. confusum were attractant to adults of both
species, but repellent to T. confusum larvae. T. castaneum larvae were attracted to
flour conditioned by T. confusum whilst repelled by flour conditioned by the
conspecies. T. confusum was more responsive than T. castaneum, but there was no
significant difference (P > 0.05) in response between sexes in both species.
INTRODUCTION
A medium in which Tribolium beetles have lived for some period is called
“conditioned” (Park, 1937; Sokoloff, 1974). A few beetles are enough to condition
the flour medium giving a persistent and disagreeable odour, turning the flour pinkish
(Chittenden, 1896; Engelhardt et al. 1956), adversely affecting the viscous and
elastic properties of the flour and create a disgusting taste (Payne, 1925).
Conditioning is one of the major stimuli for dispersal and aggregation by Tribolium
(Loconti and Roth, 1953) and it depends on the number and sex of insects, and the
duration of occupation (Ghent, 1963; Mondal, 1983, 1985). Conditioning of the
medium involves the depletion of the nutritive value of the medium; accumulation of
exuviae, dead insects and similar debris; and most markedly, accumulation of the
quinones (Roth, 1943) and pheromones (Suzuki and Sugawara, 1979) given off by
the adults Tribolium and taken up by the medium (Ghent, 1963; Sokoloff, 1972).
Flour conditioned by adult conspecifies was repellent to T. castaneum adults and
larvae, but attractive to T. confusum adults due to the production of quinones (Ghent,
1963; Ogden, 1969; Mondal, 1983, 1985). Later, adults of both sexes of T.
confusum (Ryan and O’Ceallachain, 1976; O’Callaghan, 1977; O’Ceallachain and
Ryan, 1977; Hughes, 1982), T. castaneum (Suzuki and Sugawara, 1979) and T.
brevicornis (Faustini et al., 1982a, b) were reported to be attracted to flour
conditioned by male conspecifies which was explained as being due to the
production of an aggregation pheromone (Suzuki and Sugawara, 1979; Suzuki,
1980).
All the previous works were on the intraspecific responses to conditioned medium.
However, there is no information concerning the interspecific responses of Tribolium
species to naturally conditioned medium. This led to the present experiment.
MATERIALS AND METHODS
The experiment was conducted in a ‘Choice Chamber’ consisting of a plastic Petri
dish (9 cm diameter). A filter paper was placed on the floor of the dish and the Petri
dish was divided into two equal halves by a mark on the filter paper. Approximately
1 g of fresh flour was placed at the middle of one half of dish (control) and 1 g of
conditioned flour was placed at the middle of the other half. Twenty adults of either
sex or larvae were introduced at the middle of the dish and was left uncovered, thus
providing an option for the test insects to select either the fresh flour or conditioned
flour (Mondal, 1983). The Petri dish was kept in an incubator at 30 0c. The number
of insects invaded the conditioned flour was recorded after one hour.
The tests were replicated five times for each treatment and conducted with adults of
both sexes and larvae separately. Flour media inhabited by T. castaneum and T.
confusum populations separately for approximately nine months were used as
conditioned media. Larvae aged 16 days which correspond the sixth instar (Mondal,
1984) and adults of both sexes aged between 10-20 days were used as test insects
in the experiment. Sexes of adults were determined in the pupal stage (Halstead,
1963). In each test fresh adults or larvae were used. The conditioned media were
obtained from the Department of Agricultural and Environmental Science, University
of Newcastle upon Tyne, United Kingdom.
RESULTS AND DISCUSSION
The results of the experiments and statistical analyses are shown in Table 1. The
results were rested using chi square based on the expected distribution of 50 : 50.
The media conditioned by both T. castaneum and T. confusum were attractant of
adults of both species, but were repellent to T. confusum larvae. T. castaneum
larvae showed variable responses. They were attracted to flour conditioned by T.
confusum whilst repelled by flour conditioned by the conspecies. T. confusum were
more responsive to conditioned media than T. castaneum. There was no significant
difference (P > 0.05) in response between sexes of both species.
The attraction of medium conditioned by T. castaneum and T. confusum beetles to
adults of both species is similar to the findings of Ryan and O’Ceallachain (1976),
O’Ceallachain and Ryan (1977), O’Callaghan (1977), Suzuki and Sugawara (1979),
Faustini et al. (1982a, b), Hughes (1982) and Mondal (1985) who reported that
medium conditioned by male conspecifies was attractive to the adults of both sexes
of T. castaneum, T. confusum and T. brevicornis. It is possible that the adult
attraction to these conditioned media might be due to the presence of an
aggregation pheromone (4, 8-Dimethyldecanal) produced by males (Suzuki and
Sugawara, 1979; Suzuki, 1980). The present result also support the findings of
Ghent (1963) and Ogden (1969) with T. confusum, but contrast with T. castaneum
who reported flour conditioned by conspecific adults was repellent to T. castaneum
adults, whilst attractive to T. confusum adults.
Larvae of T. confusum were repelled by the flour conditioned by beetles of both
species. T. castaneum larvae also showed similar response to medium conditioned
by conspecies only. This repellent result is similar to those of Mondal (1983, 1985)
who reported larvae of T. castaneum were repelled by medium conditioned by adult
conspecifics at high densities or during long period. This repellent effect might be
due to the presence of the quinones and / or accumulation of exuviae, dead beetles
and similar debris as explained by Mondal (1983, 1985).
In the present experiments, the medium was conditioned for long period (9 months)
by the populations (adults, larvae and pupae) of T. castaneum and T. confusum
separately in an incubator under same conditions. Although the exact numbers of
conditioning beetles were not known, but densities of the beetles in both media were
extremely above the realistic upper limit (Ogden, 1969). Both media were heavily
conditioned turning into pinkish colour which indicated the presence of quinones
produced by both sexes of adults (Mondal, 1992). According to the reports of
previous workers the conditioned media used in the present experiments were
supposed to be repellent to adults and larvae of both species due to the presence of
quinones, but the result was quite different. They were attractant. However, this
attraction does not seem to be due to an aggregation pheromone produced by the
male adults because it is produced only at low densities of the beetles or during the
short period of conditioning. It is possible that both quinones and pheromones are
secreted at continuous rates in all conditions. However, the pheromone is volatile
and, therefore, whilst present early on, does not accumulate. The quinone, on the
other hand, accumulates and eventually masks the pheromone. When the flour was
conditioned for nine months the quinones would be dominant over the pheromone.
Quinones are pheromone parsimony (Blum, 1970).
They act as epideictic
pheromones at high population densities by dissolving aggregation pheromone
globules and thus, under stressful conditions of overcrowding and lack of sufficient
food resources, the pheromone is inhibited (Faustini and Burkholder, 1987).
The attraction of conditioned media in the present experiment may be due to the
presence of other semiochemical secreted by adults or larvae in addition to the
previously reported quinones or pheromones which is not known. It needs further
investigation. However, the present results indicate that conditioning of medium may
be due to the presence of quinones, phermones or other unknown semiochemical(s).
Table 1. Number and percentage of T. castaneum and T. confusum adults and
larvae in fresh medium (control) and conditioned medium
Conditioning
species
Responding
species
Life
stage
Distribution of beetles
Nos. in
Nos. in
% total in
fresh conditioned conditioned
T.castaneum T.castaneum Male
33
67
67.0
Female
33
67
67.0
Larvae
56
44
44.0
T. confusum Male
15
85
85.0
Female
03
97
97.0
Larvae
71
29
29.0
T. confusum T. confusum Male
07
93
93.0
Female
03
97
97.0
Larvae
70
30
30.0
T.castaneum Male
23
77
77.0
Female
21
79
79.0
Larvae
29
71
71.0
Five replicates per test, each replicate consisting of 20 insects (n = 100)
NS – Not significant, P > 0.05; * Significant, P < 0.001
X2
(1df)
11.56*
11.56*
1.44NS
49.0*
88.36*
17.64*
73.96*
88.36*
16.00*
29.16*
33.6*
17.64*
CONCLUSION
Although in the present experiment the exact chemical rendering the flour medium
conditioned is unknown, but it is important that it can be used in the control of
Tribolium populations as either attractant (Dethier et al., 1960) or repellent (Dethier,
1956; Painter, 1967). Both attraction and repulsion are based on olfactory causes,
since the beetles used in the experiments responded whilst not in contact with
conditioned medium. Bags of grains or containers treated with this chemical may
prevent Tribolium as repellent from attacking and infesting the food (Mondal and
Port, 1984a, Mondal, 1989). Alternatively, as attractant it may also be used to detect
and lure populations of Tribolium into traps where they can be easily killed or
destroyed by suitable insecticides, pathogens or mechanical method (Mondal and
Port, 1984b).
ACKNOWLEDGEMENTS
The author is indebted to Dr. G.R. Port for his suggestion during the work. The
author wishes to thank the British Council for financial support under Newcastle and
Rajshahi Bilateral Link Programme and the University of Rajshahi, Bangladesh for
granting study leave.
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Blum, M.S. 1970. The chemical basis of insect sociality. In : Chemicals Controlling
Insect Behavior (Ed. M. Beroza). Academic Press, New York : 69-94.
Chittenden, F.H., 1896. Insects affecting cereals and other dry vegetable foods.
U.S. Dept. Agric. Divn. Ent. Bull. 44 : 112-131.
Dethier, V.G. 1956. Repellents. Ann. Rev. Ent. 1 : 181-202.
Dethier, V.G., Browne, L.B. and Smith, C.N. 1960. The designation of chemicals in
terms of the responses they elicit from insects. J. econ. Ent. 53 : 34-136.
Engelhardt, M., Rapoport, H. and Sokoloff, A. 1965. Odorous secretions of normal
and mutant Tribolium confusum. Science 150 : 632-633.
Faustini, D.L. and Burkholder, W.E. 1987.
Quinone-aggregation pheromone
interaction in the red flour beetle. Anim. Behav. 35 : 601-603.
Faustini, D.L., Rowe, J.R. and Burkholder, W.E. 1982a.
A male-produced
aggregation pheromone in Tribolium brevicornis (Leconte) (Coleoptera :
Tenebrionidae) and interspecific responses of several Tribolium species. J.
stored Prod. Res. 18 : 153-158.
Faustini, D.L., Post, D.C. and Burkholder, W.E.1982b. Histology of aggregation
pheromone gland in the red flour beetle. Ann. Entom. Soc. Am., 75 : 187-190
Ghent, A.W. 1963. Studies of behaviour of Tribolium flour beetles. I. Contrasting
responses of T. castaneum and T. confusum to fresh and conditioned flours.
Ecology 44 : 269-283.
Halstead, D.G.H. 1963. External sex differences in stored products Coleoptera.
Bull. Ent. Res., 54 : 119-134.
Hughes, A.L. 1982. Attraction of adult Tribolium confusum to flour conditioned by
male conspecifics. Behav. Processes 7 : 247-253.
Loconti, J.D. and Roth, L.M. 1953. Composition of the odorous secretion of
Tribolium castaneum. Ann. Entom. Soc. Am. 46 : 281-289.
Mondal, K.A.M.S.H. 1983. Response of Tribolium castaneum larvae (Herbst) to the
different components of conditioned medium. Tribolium Inform. Bull. 23 : 104111.
Mondal, K.A.M.S.H. 1984. A method of determining the larval instars of Tribolium
castaneum Herbst (Coleoptera : Tenebrionidae). Lab. Practice 33 : 120-121.
Mondal, K.A.M.S.H. 1985. Response of T. castaneum larvae to aggregation
pheromone and quinones produced by adult conspecifics. Int. Pest Control 27
: 64-66.
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methylquinone. Bangladesh J. Zool. 17 : 165-168.
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and Prospects. Tribolium Inform. Bull. 32 : 78-90.
Mondal, K.A.M.S.H. and Port, G.R. 1984a.
Repellent effect of synthetic
methylquinone on larvae of Tribolium castaneum Herbst (Coleoptera :
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to synthetic aggregation pheromone. Ent. Exp. and appl. 36 : 43-46.
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O’Ceallachain, D.P. and Ryan, M.F. 1977.
Production and perception of
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THE SIMULATION OF SIRE GENETIC EVALUATION
USING TRIBOLIUM CASTANEUM FOR THE
COMPARISON OF TWO STATISTICAL METHODS
Zhang Lao, Wang Xibin, Zhang Qineng, Wang Yachun and Liu Wei
Department of Animal Genetics and Breeding
Beijing Agricultural University
Beijing, 100094 P.R. China
ABSTRACT
The simulation of sire genetic evaluation by using Tribolium castaneum for the
comparison of two statistical methods : Henderson’s method 3 and REML for the
estimation of variance and covariance components and also the comparison of two
selection methods. The same data set of pupal length and pupal width in two
generations was analyzed by two methods. Results showed that heritabilities of two
traits estimated by Henderson’s method 3 were lower than that estimated by REML
and did not remain stable in two generations. The ranking of each family average of
breeding value showed that the usual method may be used for sire evaluation when
the heritabilities of selected traits are middle in heigh.
INTRODUCTION
Selection for improved performance of an economic trait using a selection
index. The value of selection by indices is dependent on both the accuracy (Lin et
al. 1979) and the variance of parameters (Hill, 1974).
Henderson’s method 3 is useful for all general mixed models, unbiased and
translation invariant, not unique.
It is more difficult than method 1 or 2
computationally and is more accurate than method 1 or 2. The procedure of
disadvantage : for most models there are more reductions that can be computed
than are necessary to estimate the variances. There are no rules for selecting those
reductions that will give variance component estimates with the lowest sampling
variances.
REML (Restricted Maximum Likelihood) has general characteristics :
translation invariant, estimates required to be within the parameter space, must be
solved interactively, requires normality. REML can be derived from the MIVQUE
algorithm or by maximum likelihood.
For experimental verification of theoretical aspects of quantitative genetics,
the experimenter is interested in using an organism which has a shorter generation
interval and which is more economical to maintain, Tribolium castaneum offers many
advantages for this work.
The simulation of sire genetic evaluation by using T. castaneum for the
comparison of two statistical methods : Henderson’s method 3 and REML for
estimation of variance and covariance components and also be comparison of two
selection methods.
MATERIALS AND METHODS
1. Foundation population : WT Tribolium castaneum in Lab. Of Dept. of Animal
Genetics and Breeding, Beijing Agricultural University.
2. Medium : 95% whole wheat flour and 5% yeast.
3. Incubator : The beetles were maintained in an incubator at 320c and 60%
relative humidity.
4. Generation Zero : 30 single-pair random matings for 5 days before 24-hour
egg collection came from the foundation population. They were used as a selected
population; Four 5-pair random mating groups were used as a control population.
5. Selection experiment extended over two generations and consisted of four
replication with 215 progenies per generation per replication.
6. Two methods :
(1) Henderson’s method 3
(2) REML and BLUP
7. Models :
(1) The sire model used for this analysis was :
Where :
Yijklm : phenotypic value of trait m
µm
: overall mean of trait m (m = 1,2)
Him
: replicate – day block ejject (i = 1,2,3,4)
Sexjm : sex ejject (j = 1,2)
µkm
: sire ejject (k = 1,2,….30)
eijklm : random error term
(2) The animal model used for this analysis was the same.
Where, µkm : individual effect
8. Ranking breeding value of the sires using the two methods.
RESULTS AND DISCUSSION
1.
F tests of family, replicate-day and sex effects were significant at P < 0.01
(Table 1). It suggested that it is necessary to eliminate the fixed effects while
estimating the breeding value.
Table 1. Analysis of variance for pupal length and width of generation zero
** : Significant at P < 0.01
2.
The genetic parameters of pupal length and width were different using two
method, and the genetic parameters were equable using REML than Henderson’s
method 3 in the two generations.
Table. 2. Genetic and phenotypic parameters estimated by two methods
3.
Sire evaluation using two methods was nearly the same for the trait which has
the mid-high h2. The best five sires in the ranking of the sire breeding value were the
same using the two methods. The rest of five sires selected were 80% the same,
although the ranking was not the same.
Table 3. The sire evaluation using two selection methods
Rank
Sire number
method I
Sire number
method II
1
14
2
12
3
11
4
15
5
22
6
21
7
20
8
2
9
4
10
24
14
12
11
15
22
2
4
24
26
21
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Prediction and Variance Component Estimation. Univ. of Guelph
9. Wilton, J.W. 1984. Course Notes for Selection Principles in Animal Breeding.
Univ. of Guelph.
ACKNOWLEDGEMENTS
This research was supported by funding from the Natural Sciences Council of
China.
McGraw, Page and Alexander Sokoloff
Biology Department
California State University
San Bernardino, California 92407
The repair of genetic damage in Tribolium freemani black and T. freemani – T.
castaneum black hybrids through injection of b-alanine.
INTRODUCTION
Catecholamines such as b-alanine, dopamine, N-acetyl-dopamine and N-balanyldopamine are intricately involved in pigment and cuticle sclerotization in T.
castaneum (Kramer et al., 1987). A mutant strain of T. castaneum, T. castaneum
black has been shown to recover from genetic damage when treated with b-alanine
in the teneral adult stage (Kramer et al., 1984). Spray and Sokoloff, unpublished,
have identified a black mutant in T. freemani. T. freemani and T. castaneum are
both in the castaneum species group, and when hybridized produce abundant
progeny (Nakakita et al., 1981, Brownlee and Sokoloff, 1988). Also, autosomal
mutants in T. freemani have been found to have superficial resemblance to known
mutants of T. castaneum (Carrillo and Sokoloff 1991). The above, along with the
identification of homologous genes in T. castaneum and T. freemani (Spray and
Sokoloff 1991), give support to the relative closeness of the two species.
This experiment was to determine if injection of b-alanine into T. freemani black
would produce the wild-type body color and also to determine if hybrids between T.
castaneum and T. freemani black would produce the wild type color when injected
with b-alanine. This study gives support to the idea that the species have very
similar if not identical biochemical pathways leading to the production of body color
and cuticle sclerotization.
MATERIALS AND METHODS
Three Tribolium stocks were used in this study. (1) A pure breeding strain of T.
castaneum black. (2) A pure breeding strain of T. freemani black. (3) A cross of T.
freemani black and T. castaneum black for the purpose of obtaining hybrids. The
medium was prepared from approximately 5 grams of whole wheat flour containing
0.02 grams of Fumidil-B (Bicyclohexylammonium Fumagillin, an anti-parasitic drug).
All stocks were kept in an incubator at 290c and 70 percent r.h. To assure abundant
progeny, the parental generation was transferred weekly to a new set of creamers
containing a fresh medium. After a period of 3 weeks, the emerging pupae, ranging
from opaque to black in eye color, were removed from the creamers. A 33 gage
micro-syringe was used to inject approximately 0.1 micro liters of a 4 molar b-alanine
solution. This dosage had proven optimal in previous experimentations (Roseland et
al., 1983). Each pupa was then immobilized using forceps. The needle was then
inserted between the third and fourth segments directed towards the thorax. The
wounded pupae were then placed in a bed of flour and allowed to recover and
proceed in their development in the incubator. After a 2 to 3 day period, the beetles
were removed and examined according to pigmentation.
Notes-Research, Teaching and Technical
RESULTS AND DISCUSSION
The expression of wild-type pigmentation from the T. freemani black and the T.
freemani – T. castaneum hybrids was successful. However, we were not able to
recover any of the T. castaneum black that were injected. Of the 248 T. freemani
black pupae injected with @ 0.1 micro liters of 4 molar b-alanine solution, only 6
percent developed the wild-type color. Moreover, of the 50 T. freemani – T.
castaneum black hybrids, 14 percent became chestnut. The other pupae did not
survive the injury and / or received too much or not enough b-alanine to emerge as
chestnut. Our crude instrumentation also played a part in the numbers. However,
as time progressed, we became more skilled. The results are listed in Table 1.
Our project differed from Kramer et al., 1984 in that we chose to use the pupal stage
for the injection of b-alanine. This was for several reasons : (1) The pupal stage
allowed a broader time range within which injections could be accomplished. (2) The
pupae were much easier to secure than the teneral adult stage. (3) The levels of
catecholamines were the same for the wild-type and the black strains at this stage
(Kramer et al., 1984).
These results, along with the fact that the two species are able to produce hybrids of
mutants demonstrated to have homologous genes, lead to the idea that the species
have only recently diverged and that they may have retained similar or if not identical
pathways in the production of body color and cuticle sclerotization.
Literature Cited
Brownlee, A. and A. Sokoloff. 1988. Transmission of Tribolium castaneum (Herbst)
mutants to T. castaneum – T. freemani Hinton hybrids (Coleoptera : Tenebrionidae).
J. Stored. Prod. Res. 24 : 145-50.
Carrillo, L. and A. Sokoloff. 1991. New mutants and teratologies in Tribolium
freemani Hinton. Tribolium Inform. Bull. 31 : 55-60.
Kramer, K.J., Morgan, J.D., Hopkins, T.L., Roseland, C.R., Aso, Y., Beeman, R.W.,
and Lookhart, G.L. 1984. Catecholamines and b-alanine in the red flour beetle
Tribolium castaneum. Roles in cuticle sclerotization and melanization. Insect
Biochem. 14 : 293-8.
Kramer, K.J., Roseland, C.R., Hopkins, T.L. 1987.
Cuticular strength and
pigmentation of rust-red and black strains of Tribolium castaneum – correlation with
catecholamine and b-alanine content. Insect Biochem. 17 : 21-8.
Nakakita, H., Imura, O., and Winks, R.G. 1981. Hybridization between Tribolium
freemani Hinton and T. castaneum (Herbst), and some preliminary studies on the
biology of T. freemani (Coleoptera : Tenebrionidae). Appl. Ent. Zool. 16 : 209-15.
Roseland, C.R., Beeman, R.W., Kramer, K.J., and Hopkins, T.L. 1983.
Microinjection of amino acids in Tribolium. Tribolium Inform. Bull. 23 : 132.
Table 1. Numbers in relation to survival and pigmentation
T. freemani
T. castaneum
T. c. – T. f.
Injected
247
25
50
Survived
66
0
30
Chestnut
16
7
Black
49
23
Shengfang, Z. and Yongping, L.
Institute of Plant Quarantine
Ministry of Agriculture
Beijing 100026, China
* Identification of four species of the castaneum species-group of Tribolium
(Coleoptera : Tenebrionidae).
Tribolium freemani Hinton was rediscovered in 1981 in Japan from imported corn.
We believe that the species of Tribolium in China previously described as T. madens
actually should be T. freemani by examining intercepted specimens sent by Dr.
Nakakita. The species is widely distributed in the Sinkiang Province. Besides, it was
rarely found in the following provinces of China : Gansu, Ninzin, Shanxi and Yunnan.
It causes some damage to several stored products such as wheat, wheat bran and
wheat flour, corn and corn flour, rice apricot stones. Several living adult and larva
specimens also were collected from dry ground beetles and tortoise shells in drug
stores.
Halstead (1969) gave characters to distinguish T. audax and T. madens. This paper
provides comparisons of T. audax, T. madens, T. freemani and T. castaneum (Table
1). The data of T. audax and T. madens is based on Halstead’s studies.
By comparing male genitalia of four species (Figs. 1-12) it is revealed that the
genitalia of T. audax is much more similar to that of T. madens that to that of T.
freemani or T. castaneum, and the genitalia of T. freemani is much more similar to
that of T. castaneum than to that of T. audax and T. madens.
Crosses made between T. audax and T. madens produced a few infertile F1 hybrids
(Halstead, 1969). Interspecific crosses between T. freemani and T. castaneum
produced many infertile F1 hybrids. Crosses between T. freemani and T. madens
and between T. freemani and T. audax were tried, but no F1 progey were obtained.
So the conclusion is that, in that castaneum-section, T. freemani is closer to T.
castaneum than to T. madens or T. audax (Nakakita, 1983). The conclusion is
supported morphologically by comparing the genitalia of the four species.
References
(1)
(2)
(3)
(4)
Hinton, H.E. (1984). A synopsis of the genus Tribolium Macleay, with some
remarks on the evolution of its species-group (Coleoptera : Tenebrionidae).
Bull. Ent. Res., 39 :13-56.
Halstead, D.G.H. (1969). A new species of Tribolium from North America
previously confused with Tribolium madens (Charp.) (Coleoptera :
Tenebrionidae). J. stored Prod. Res., 4 : 295-304.
Nakakita, H. et al. (1981). Hybridization between Tribolium freemani Hinton
and Tribolium castaneum (Herbst) and some preliminary studies on the
biology of T. freemani (Coleoptera : Tenebrionidae). Appl. Ent. Zool. 16 : 209215.
Nakakita, H. (1983). Rediscovery of Tribolium freemani Hinton a stored
product insect unexposed to entomologists for the past 100 years. JARQ 16 :
239-245.
Sokoloff, A.
Biology Department
California State University
San Bernardino, California 92407
* A succession of habitats in laboratory cultures.
As pointed out by Lezcano and Sokoloff elsewhere in this issue of TIB, overcrowding
either as an incidental occurrence during the course of an experiment or deliberate,
involving last instar larvae only, adults only or all stages of flour beetles, may result
in drastic changes in the flour medium, leading to the loss of the culture.
The changes are associated with an increase of moisture within the culture resulting
from the metabolic activities of the beetles. Moisture forms in tiny droplets on the
inside surface of the vial or any glass container, and it is gradually absorbed by the
flour. The flour then gradually loses its particulate nature and becomes sticky. At a
certain point, mold spores are stimulated to form mycelia and as the mat the mycelia
form becomes thick, it will entrap some of the larvae or adults. The survivors are
driven to the surface of the medium. Starvation may force the beetles back into the
sticky medium. As moisture is reduced, the sticky flour may be covered with fungal
mycelia which then can serve as food for the flour beetles.
As the flour dries, it forms a plug which detaches itself from the container’s walls, or
it may turn to a black liquid in which the beetles drown.
Gradually the beetle population decreases, and as the plug dries further the
survivors may use it as a source of food. Some larvae may move toward the surface
of the plug, or to the galleries which both the larvae and adults have dug within the
plug. The larvae may pupate in the tunnels and complete their development,
producing more adults, but usually the population within the container declines and
dies.
The changes have been observed to occur in one dram vials or in pint milk glass
bottles. The essential factor is to have a dense population (for Tribolium freemani
the critical density appears to be about 100 last instar larvae or 100 adults per gram).
During an experiment requiring a great deal of replication, the larvae, pupae and
adults of Tribolium freemani were scored, the adults were discarded into a milk bottle
and the larvae were placed in another bottle. Both bottles were about ¾ filled with
unbleached wheat flour, ad remained unplugged during these observations. As the
density of larvae or adults increased in these bottles the medium underwent the
changes described above and in Lezcano and Sokoloff’s paper. The flour in the
bottle containing larvae soon became very moist and there were droplets of moisture
developing on the walls of the bottle. The medium became moldy, and as the
medium started to dry it formed a plug which gradually became detached from the
inner walls of the glass bottle. The beetle larvae were either driven to the surface or
were trapped in the plug.
At this point I noted that there were some Diptera larvae crawling on the surface of
the medium. Some adult flies were seen going in and out of the uncapped bottle.
They proved to be humpbacked flies (Family Phoridae), which previous experience
had shown that they can be easily raised to a culture in Drosophila medium. The
files run in jerky movements, and they seem pretty aggressive, often landing on
one’s face and hands. According to Borror, Triplehorn and Johnson, 1989 “Phorids
are small or minute flies that are easily recognized by their humpbacked
appearance, characteristic venation, and the laterally flattened hind femora. The
adults are fairly common in many habitats, but most abundant about decaying
vegetation. The habits of the larvae are varied : some occur in decaying animal or
vegetable matter; some occur in fungi; some are internal parasites of various other
insects; and some occur as parasites or comensals in the nests of ants or termites.
A few of the species that occur in ant or termite nests (and some others as well)
have the wings reduced or lacking. More than 350 species occur in our area.
When phorid larvae were noticed, the bottle was plugged. The larvae pupated, and
eventually about 100 adults emerged, making it possible to identify this fly readily to
the family Phoridae.
Although such successional changes of one organism to another one the same
medium are artificial, nevertheless, as these observations show, it is possible for
organisms whose larvae live in a xeric habitat (in the present case Tribolium
freemani) to be succeeded by organisms whose larval habitat is mesic (flies of the
family Phoridae).
It is doubtful that these two organisms, Tribolium and the hump-backed flies, would
ever come into an association under normal circumstances. However, Tribolium has
been reported to consume the flesh of dead birds or insects, and phorids appear to
be opportunistic scavengers. So, if the opportunity presented itself, flour beetles and
flies may enter into a competitive relationship.